Introduction：Neonatal tetanus is a major cause of neonatal mortality in many developing countries and remains a major public health problem. This study aimed to determine risk factors associated with neonatal tetanus in Wenzhou, China.Methodology：Medical records of neonatal tetanus cases from 17 hospitals over a 13-year period (2000–2012) were reviewed for potential risk factors. Controls were selected from neonates with diseases other than tetanus who were admitted to the same facility during the same period. The potential risk factors of the neonatal tetanus group were compared with the control group using univariate analysis and an unconditional logistic regression model.Results：A total of 246 neonates with tetanus and 257 controls were included in this study. Univariate analysis showed that having untrained birth attendants, home delivery, an unsterile method of delivery and being a migrant to Wenzhou were significantly different between the two groups (P < 0.001). Logistic regression analysis revealed that the odds of having an untrained birth attendant, home delivery and an unsterile method of delivery were significantly higher in the tetanus group than the control group (odds ratio: 1371.0; 95% confidence interval: 206.0, 9123.5).Conclusion：This study identified that the main risks of neonatal tetanus in cases from Wenzhou were having an untrained birth attendant, home delivery and an unsterile method of delivery. Preventive measures directed to these risk factors may reduce the occurrence of neonatal tetanus in the studied area.

OBJECTIVETo analyze the dental care utilization and expenditure of residents in Beijing, and to provide some basis on the policy of oral health insurance system.

METHODSA cross-sectional survey was conducted among 1,517 subjects (urban area) and 1,878 subjects (rural area) of all age groups in Beijing selected by stratified, clustering, random sampling. The data of oral health care utilization and expenditure were collected in their home.

RESULTSThe number of the people who visited a dentist in a year were low both in urban area and in rural area, but the expenditure for oral health care per visit were quite high. The value of utilization of dental care in rural residents was 1/3 of that in urban residents, while the value of expenditure in rural people was about 1/2 of that in urban people. 2.07% incomes of rural residents were used for dental care per year, the corresponding value of urban residents was 1.77%. There was significant difference on the expenditure among those with different demographic, socio-economic backgrounds.

CONCLUSIONSThe expenditure for oral health care was high in Beijing, which accounted for quite a lot in average incomes per year. The burden of expenditure for dental care on rural residents was heavier than that on urban residents. The level of expenditure for dental care could provide some references for oral health insurance system in Beijing.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; Dental Health Services ; economics ; statistics & numerical data ; Fees, Dental ; Female ; Health Services Accessibility ; statistics & numerical data ; Humans ; Infant ; Infant, Newborn ; Insurance, Dental ; Male ; Middle Aged ; Rural Health Services ; statistics & numerical data ; Urban Health Services ; statistics & numerical data

This study is to establish the characteristic HPLC chromatogram of phenols in Ephedrae Herba, from which to pick out the marker peaks, followed by the analysis of the regularity of their distribution and content in the herbaceous stems of Ephedra sinica, E. intermedia and E. equisetina. The HPLC-DAD method for the characteristic chromatogram as well as quantitative analysis was established. The separation was carried out on a YMC-Pack ODS-A column (4.6 mm x 250 mm, 5 µm), eluted with the mobile phases as 0.01% formic acid aqueous solution (A) and acetonitrile (B) in a linear gradient (0-10 min, 17% B; 10-25 min, 17%-19% B; 25- 33 min, 19%-48% B; 33-35 min, 48%-51% B; 35-44 min, 51% B). The flow rate was kept at 1.0 mL · min⁻¹. The column tem- perature was 40 °C, and the detection wavelength was set at 350 nm (0-16 min) and 330 nm (16-44 min). Forty-six batches of collected samples from three official origins of Ephedrae Herba were detected, whose liquid chromatograms proven to be helpful to the differentiation of different origins. With principal component analysis and the analysis of distribution of peak area, twelve key peaks from the chromatogram were discussed in details on their contributions to the characteristics and differences of three official origins of the herb: peak area of peak 10, 11, 12 were found out to be significantly higher in E. equisetina than in other two origins, whose sum (higher than 146 mAU in E. equisetina) was useful for the discrimination between E. equisetina and the other two origins; peak area of 1 and 4 were respectively higher in E. sinica and E. intermedia than in other official origins, indicating their important effect on the differen- tiation of corresponding origins; peak 8 and 9 were picked out as two characteristic common peaks in three official origins of the herb, whose peak area showed little difference among different origins; further, peak area of other key peaks in the chromatogram also showed some difference among three origins, which make contributions to the differentiation of origins as well. Then, four phenols as 2"-O-α- L-rhamnosyl-isovitexin (1), vitexin (2), pollenitin B (5) and herbacetin-7-O-β-D-glucoside (6) were quantitative analyzed with the above-mentioned method, with good linear relationship and accuracy (recoveries in a range of 97.8%-102.5%). The content of the four phenols were firstly reported in Ephedrae Herba from official origins, which were respectively trace-1.55 (1), trace-0.160 (2), trace-0.284 (5) and trace-0.620 (6) mg · g⁻¹ in all of the tested samples. In addition, the content of these phenols showed differences in three official origins, especially 1, whose content in E. sinica [(0.670 ± 0.88) mg ± g⁻¹] were significantly higher than in other two origins (lower than 0.16 mg ± g⁻¹ besides sample Ei-060630-2-2), and 6, whose average content in E. equisetina [(0.260 ± 0.039 2) mg · g⁻¹] were twice as high as in E. sinica [(0.120 ± 0.270) mg · g⁻¹] and E. intermedia [(0.136 ± 0.485) mg g⁻¹], indicating the important effects of the two constituents on the differentiation among three official origins of the herb. The method established for the characteristic HPLC chromatogram and quantitative analysis of phenols was simple and accurate, and the marker constituents selected may provide new guides for the discrimination of official origins as well as the improvement of quality criteria of EphedraeHerba.
Chromatography, High Pressure Liquid ; methods ; Ephedra ; chemistry ; Phenols ; analysis

Objective:To explore and optimize the teaching content of clinical case discussion course based on clinical pathophysiology and therapy (CPPT), and to improve the teaching quality of this course.Methods:Self-compiled evaluation form was sent to all the students who participated in this course in 2016-2017 school year, and the teaching satisfaction and teaching characteristics were evaluated. The differences of evaluation results from students with different educational programs or teachers with different seniority were compared, and the correlation between evaluation results and teachers' seniority was analyzed.Results:The students had a high evaluation on the course, but students from different programs had significant different comments on thinking inspiring, speech encouraging, questions guiding, process holding and effective feedback, as well as the self-evaluation of mastery degree. For teachers with different seniority, there were significant differences in scores in all aspects except speech encouraging, but no differences in mastery degree. Spearman analysis showed that there was a certain correlation between scores and teachers' seniority.Conclusion:The overall situation of the CPPT case discussion course in our school is worthy of recognition, but it needs to be improved in many aspects.

OBJECTIVETo investigate the expression of osteopontin mRNA and its correlation with clinicopathologic features of gastric cancer and elucidate its role in tumor invasion and distant metastasis.

METHODSThe expression of OPN mRNA was detected by semi-quantitive RT-PCR. The relationship between the relative content of OPN mRNA and clinicopathologic features of gastric cancer was analyzed.

RESULTSIn 66 cancer tissue samples, a 330 bp band was detected in 50 cases, the positive rate of OPN mRNA expression was 75.8% (50/66). The expression in all 20 cases of normal gastric mucosa was negative. The expression was associated with the depth of tumor invasion, diameter, lymph node metastasis and but had no correlation with differentiation grades. The 66 patients were followed up for 10 approximately 27 months (mean 16 months). The OPN mRNA expression positive group (50 cases) had recurrence in 15 patients and the negative group (16 cases) had only 1 case with recurrence (P = 0.05); 10 patients died in OPN mRNA expression positive group but no patient died in OPN staining negative group (P = 0.05).

CONCLUSIONOPN mRNA is over-expressed in gastric cancer. It reflects the progression of disease and association with poor prognosis of gastric cancer. OPN may play an important role in the process of distant metastasis in gastric cancer.

Adenocarcinoma ; metabolism ; pathology ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Osteopontin ; biosynthesis ; genetics ; Prognosis ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach Neoplasms ; metabolism ; pathology

Objective To explore the optimal experiment conditions of CCK-8 and MTS for cell proliferation assays in human amniotic epithelial cells and to evaluate the cytotoxicity of these reagents. Methods Human amniotic epithelial cells (hAECs) in logarithm growth stages were prepared in different cell concentrations with DMEM/F12 and 10% FBS. The sensitivity and optimal wavelengths was determined based on the optical density (OD) measured at 450 nm and 492 nm. The optimal time was determined under the conditions of the same cell concentration and defined OD values. HAECs were treated with DMSO, CCK-8 and MTS for 1 h, 2 h, 3 h, and 4 h, respectively. 24 h later, cytotoxicity of the CCK-8 and MTS was evaluated by determination of cell proliferation and Trypan Blue staining. Results The optimal detection wavelength was 450 nm for CCK-8, and 492 nm for MTS. The sensitivity of CCK-8 was slightly lower then that of MTS. The optimal time for incubation hAECs with CCK-8 was 4 h within 1~4 h. The inhibitory on cell proliferation and cytotoxicity of CCK-8 were weaker then those of MTS. Conclusion CCK-8 is a convenient reagent with low cytotoxicity for detection of the proliferation of hAECs.

OBJECTIVETo explore the relationship between methyl-tetra-hydrofolic acid (MTHFR) 677 gene polymorphism and the risk of stomach cancer.

METHODSA population based case-control study was conducted and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to detect its genotypes.

RESULTSAmong cases with stomach cancer, the frequency of C/C, C/T, T/T genotype were 25.8%, 54.6%, 19.6%, compared with controls as 34.5%, 50.9%, 14.6% respectively. Using C/C genotype as reference, the OR of C/T or T/T genotype was 1.52 (95% CI: 1.04 - 2.23). 53.3% C and 46.7% T allele were distributed in stomach cancer cases, while 60.0% C and 40.0% T in controls. The OR for T allele in relation to C allele was 1.31 (1.02 - 1.69) when C allele was used as reference. In addition, the present study showed that MTHFR677 AnyT genotype might interact with smoking, moldy food intake, wheat porridge intake, eating salty food and Hp CagA infection to increase the risk of stomach cancer. No interaction was observed between MTHFR677 AnyT genotype and alcohol drinking or green tea intake.

CONCLUSIONMTHFR677 AnyT genotype, might increase the risk of stomach cancer development and the genotype might also interact with other environmental risk factors to increase the risk of stomach cancer.

Adult ; Alleles ; Case-Control Studies ; China ; epidemiology ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Life Style ; Male ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Point Mutation ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Risk Factors ; Smoking ; adverse effects ; Stomach Neoplasms ; enzymology ; etiology ; genetics

OBJECTIVESevere newborn hypoxic-ischemic encephalopathy (HIE) has a very high rate of disability and no effective treatment is available. The present study aimed to preliminarily evaluate the effects of human neural stem cell transplantation in treatment of severe neonatal HIE.

METHODSThe patient was a 75-day old male infant with sequelae of severe HIE who had highly delayed development of intelligence and movement and myotonia. MRI showed multiple cerebromalacia and encephalatrophy. Cells obtained from the forebrain of an 11-week old fetus were cultured and amplified for 15 days. And then the human fetal neural stem cells were injected into cerebral ventricle of this infant.

RESULTSTwenty eight days after transplantation, remarkable improvement occurred not only in his myotonia but also in his intelligence and movement, which became similar to those of the normal infants of the same age. Positron emission tomography (PET) showed significantly increased radioactivity at temporal and occipital lobes which suggested that the cellular metabolism had increased greatly.

CONCLUSIONThe short-term effect of NSCs transplantation on the infant with severe HIE sequelae was significant. PET suggested that the implanted NSCs survived. Many more studies are needed to evaluate long-term effects of NSC transplantation in treatment of HIE.

Asphyxia Neonatorum ; complications ; Brain ; pathology ; physiopathology ; Female ; Humans ; Hypoxia-Ischemia, Brain ; etiology ; pathology ; physiopathology ; therapy ; Infant ; Infant, Newborn ; Injections, Intraventricular ; Multipotent Stem Cells ; transplantation ; Neurons ; Positron-Emission Tomography ; Prognosis ; Stem Cell Transplantation ; methods ; Time Factors ; Treatment Outcome

OBJECTIVENeonatal hypoxic-ischemic encephalopathy (HIE) harms the lives and health of newborn infants and children severely. Given the absence of effective therapies for HIE, it is important to derive new strategies. Neural stem cells (NSCs) have great potential as a therapeutic tool for the repair of a number of central nervous system disorders that involve cell loss. This study was designed to transplant the neural stem cells derived from human fetal brain (hNSCs) into cerebral ventricle of neonatal rat following hypoxic-ischemic injury and to investigate their survival, migration and differentiation in rat brain.

METHODSCells obtained from the forebrain of a 12-week old fetus were cultured in the presence of epidermal growth factor, basic fibroblast growth factor and leukemia inhibitory factor for 11 days. Animal models were built in 7-day-postnatal Wistar rats, 3-days after hypoxia-ischemia (HI), 5 microl suspension containing 5.0 x 10(5) hNSCs was injected into the left cerebral ventricle of each HIE rat by using stereotactic instrument. No immunosuppression therapy was given to the animals. At 1, 2, 4 weeks and 3 months after transplantation, the rats were sacrificed and brain tissues were harvested and were then examined by H-E staining and immunohistochemical analysis.

RESULTSImplanted cells expressing human nuclear protein (hNP) migrated form the subventricular zone (SVZ) along corpus callosum to the damaged areas, especially to the injured side of cortex and hippocampus. In different areas, the implanted hNSCs differentiated into different cell types which were similar to the host cells. The 85% implanted cells in cortex consisted of hNuc-NF or hNuc-Tublin double positive cells, while in the migratory way, 60% implanted cells differentiated into hNuc-GFAP double positive cells. Compared with the 1-week time point, an increased number of hNP-positive cells were observed at 2-weeks, but the number of these cells greatly decreased at 4-weeks and 3 months.

CONCLUSIONThe implanted hNSCs could extensively survive, migrate in the brain of neonatal rat with HIE and could differentiate into neurons and astrocytes in a regionally specific manner.

Animals ; Animals, Newborn ; Brain ; pathology ; Carotid Artery, Common ; surgery ; Cell Differentiation ; Cell Movement ; Disease Models, Animal ; Fetal Stem Cells ; transplantation ; Humans ; Hypoxia ; complications ; physiopathology ; Hypoxia-Ischemia, Brain ; pathology ; physiopathology ; therapy ; Immunohistochemistry ; Injections, Intraventricular ; methods ; Ligation ; methods ; Neurons ; Nuclear Proteins ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stem Cell Transplantation ; methods ; Survival Analysis ; Time Factors

The study was designed to explore the drug-drug interactions mechanisms mediated by OATP1B1 between traditional Chinese medicine Danshensu and rosuvastatin. First, the changes of rosuvastatin pharmacokinetics were investigated in presence of Danshensu in rats. Then, the primary rat hepatocytes model was established to explore the effects of Danshensu on the uptake of rosuvastatin by hepatocytes. Finally, HEK293T cells with overexpression of OATP1B1*a and OATP1B1*5 were established using a lentiviral delivery system to explore the effects of Danshensu on the uptake of rosuvastatin. Rosuvastatin pharmacokinetic parameters of C(max0, AUCO(0-t), AUC(0-∞) were increased about 123%, 194% and 195%, by Danshensu in rats, while the CL z/F value was decreased by 60%. Uptake of rosuvastatin in the primary rat hepatocytes was decreased by 3.13%, 41.15% and 74.62%, respectively in the presence of 20, 40 and 80 μmol x L(-1) Danshensu. The IC50 parameters was (53.04 ± 2.43) μmol x L(-1). The inhibitory effect of Danshensu on OATP1B1 mediated transport of rosuvastatin was related to the OATP1B1 gene type. In OATP1B1*5-HEK293T mutant cells, transport of rosuvastatin were reduced by (39.11 ± 4.94)% and (63.61 ± 3.94)%, respectively, by Danshensu at 1 and 10 μmol x L(-1). While transport of rosuvastatin was reduced by (8.22 ± 2.40)% and (11.56 ± 3.04)% and in OATP1B1*1a cells, respectively. Danshensu significantly altered the pharmacokinetics of rosuvastatin in rats, which was related to competitive inhibition of transport by OATPJBI. Danshensu exhibited a significant activity in the inhibition of rosuvastatin transport by OATP1B1*5-HEK293T, but not by OATP1B1*1a, suggesting a dependence on OATP1B1 sequence.
Animals ; Drug Interactions ; Drugs, Chinese Herbal ; pharmacology ; HEK293 Cells ; Hepatocytes ; drug effects ; metabolism ; Humans ; Lactates ; pharmacology ; Organic Anion Transporters ; metabolism ; Rats ; Rosuvastatin Calcium ; pharmacology ; Solute Carrier Organic Anion Transporter Family Member 1b1